Chromosome analysis by fluorescence in situ hybridization of callus-derived regenerants in Allium cyaneum R

• Published in: Plant cell reports Vol. 18; no. 3/4; pp. 209 - 213
• Main Authors: Lee, S.H, Ryu, J.A, Do, G.S, Seo, B.B, Pak, J.H, Kim, I.S, Song, S.D
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.12.1998; Springer Nature B.V
• Subjects: 2,4-D; Agronomy. Soil science and plant productions; Allium; Allium cyaneum; Autotetraploid; autotetraploid regenerants; benzyladenine; Biological and medical sciences; Biotin; Bulbs; Callus; Cell culture; chromosome 4; Chromosome 5; chromosome 7; Chromosome B; chromosome mapping; Chromosomes; culture media; cytogenetic analysis; Cytogenetics; digoxigenin; Diploids; dose response; Explants; Fluorescence; Fluorescence in situ hybridization; Fluorescent indicators; Fundamental and applied biological sciences. Psychology; Gene families; Generalities. Genetics. Plant material; Genes; Genetics; Genetics and breeding of economic plants; Hybridization; in vitro culture; kinetin; localization; murashige and skoog medium; naphthaleneacetic acid; Ploidy; regenerative ability; ribosomal RNA; rRNA; rRNA 18S; rRNA 26S; rRNA 5S; Satellites; somaclonal variation; somatic hybridization; Monocotyledones; Genetic variant; Fluorescence; Micropropagation; Hybridization; Somaclonal variation; Tetraploidy; Regeneration; Liliaceae; Angiospermae; Cytogenetics; Spermatophyta; Karyotype; Key words Somaclonal variation; Autotetraploid regenerants; Allium cyaneum; Fluorescence in situ hybridization
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s002990050558

Investigations were performed to confirm the optimal in vitro culture condition for callus induction and plant regeneration, to observe if somoclonal variation occurs among regenerated plants at the ploidy level and to analyse the chromosomal location of 5S and 18S-26S rRNA gene families using fluorescence in situ hybridization in callus-derived plants of Allium cyaneum. Highest callus initiation was achieved with bulb explants cultured on MS medium supplemented with 2,4-D and BAP at 1 mg l(-1) each. A total of 195 plants was obtained when using MS medium supplemented with 1 mg l(-1) NAA and 5 mg l(-1) BAP; about 92% were diploid having 2 n = 16; 8% showed a variation in ploidy level. Using digoxigenin-labelled 5S rRNA and biotin-labelled 18S-26S rRNA gene probes, we compared the fluorescence in situ hybridization patterns of autotetraploid plants with the A. cyaneum wild type. The 5S rRNA gene sites were detected on the interstitial region in the short arm of chromosome 4 and on the interstitial region in both arms of chromosome 7. The 18S-26S rRNA gene sites were detected on the terminal region of the short arm, including the satellite of chromosome 5, as well as on a part of chromosome B. The chromosomal location of both rRNA genes in regenerated autotetraploid plants corresponded to those of the wild species.